The broad and long-term objectives of this application are to develop a proteomics based lysate microarray technology for the detection of protein markers from very low amounts of samples. The multiplexing and miniaturization capability of the microarray platform allows thousands of samples to be screened in a single experiment with high sensitivity. In Year I of this application, the various steps involved in the manufacture and processing of lysate microarray technology will be optimized for detection of disease markers from cell lines commonly used in in-vitro studies of breast cancer and atherosclerosis. The disease specific protein markers that would be screened are estrogen receptor and HER-2 proteins for breast cancer and apolipoproteins -A and -B100 for atherosclerosis. The methodology will be refined to obtain highest sensitivity, reproducibility and quantification of disease markers. In Year II, the development process will be extended and applied to screening of disease markers from breast cancer and atherosclerotic patients samples obtained from ongoing clinical studies. The methodology will be validated by comparing the results generated to those obtained by standard immunological methods such as ELISA. The lysate microarray technology proposed here will enable screening of a large number of clinical samples from any disease in a highly quantifiable, reproducible and economical manner. This high-throughput technique will have a great impact on the screening of disease markers from clinical samples for the early detection of cancer and other ailments.